Dissecting inflammasome anatomy: mechanistic studies and potential intervention

描述（由申请人提供）：本提案的主要目的是阐明多蛋白复合物炎症小体的结构、激活和调节机制，并开发具有未来治疗干预潜力的干扰炎症小体组装的工具化合物。钱银博士目前是波士顿儿童医院和哈佛医学院的研究员，她的近期目标是在获得终身教职之前完成一年的指导研究。尹博士的长期目标包括独立研究人类先天免疫和炎症系统及其与病原体、环境和内在压力的相互作用，希望找到免疫和炎症疾病的合适治疗靶点和有效手段。Yin博士是一位训练有素的结构生物学家，专门研究先天免疫和炎症信号。该项目将在波士顿儿童医院进行，该医院位于波士顿朗伍德医疗区丰富的研究社区，候选人可以从邻近的哈佛医学院获得所有设施和资源。尹博士将由吴昊博士指导结构和生物物理方法来阐明炎性体结构，并由Timothy Mitchison博士共同指导高通量分析和工具化合物的开发。在一年的指导期内，尹博士将在导师的指导下学习电子显微镜和高通量筛选等新技术，并通过参加课程、研讨会和工作坊来培养专业技能。之后，尹博士计划在拟议奖项的帮助下过渡到独立。炎性小体是一种多蛋白复合物，可引发一线炎症反应。在指导阶段，Yin博士建议重建AIM2炎性体的高分辨率电子显微镜（EM）结构，并用x射线晶体学提供详细的界面相互作用（Aim 1）。本期尹博士还将阐明天然AIM2炎性体抑制剂p202的作用机制。提出的工作主要建立在ASC PYD结构域体外重构的初步EM数据上，ASC PYD结构域是大多数炎症小体的常见适配器。在独立阶段，Yin博士将1)研究NLRP3炎症小体的组装机制，阐明不同炎症小体之间的差异需求（目标2）；2)检测基于结构的稳定螺旋肽，筛选化学文库，寻找干扰炎症小体组装的小分子，以期找到具有潜在治疗价值的可药物界面和工具化合物（目标3）。

DESCRIPTION (provided by applicant): The principal aim of this proposal is to elucidate the structures and activation and regulation mechanisms of the multi-protein complex inflammasomes, and to develop tool compounds that perturb the assembly of the inflammasomes with potentials for future therapeutic interventions. Dr. Qian Yin is currently a research fellow in Boston Children's Hospital and Harvard Medical School with an immediate goal of completing one-year mentored research before establishing herself in a tenure-track academic position. Dr. Yin's long-term goals include independent researches to understand human innate immune and inflammatory systems and their interaction with pathogens and environmental and intrinsic stresses, with the hope of finding amenable therapeutic targets and effective means for immune and inflammatory diseases. Dr. Yin is a well-trained structural biologist specialized in innate immune and inflammatory signaling. The project will be conducted at Boston Children's Hospital in the rich research community of Longwood Medical Area of Boston, with all facilities and resources from neighboring Harvard Medical School available to the candidate. Dr. Yin will be mentored by Dr. Hao Wu in structural and biophysical approaches to elucidate inflammasome structures and co-mentored by Dr. Timothy Mitchison in development of high throughput assays and tool compounds. During the one-year mentored period, Dr. Yin will learn new techniques such as electron microscopy and high throughput screening under the guidance of the mentors, and develop professional skills by attending classes, seminars, and workshops. After that Dr. Yin plans to make the transition to independence with the assistance of the proposed awards. Inflammasomes are multi-protein complexes that launch the first-line inflammatory responses. During the mentored phase Dr. Yin proposes to reconstruct high-resolution electron microscopy (EM) structure of the AIM2 inflammasome, with X-ray crystallography providing detailed interfacial interactions (Aim 1). This period will also see Dr. Yin clarify the working mechanism of p202, a natural AIM2 inflammasome inhibitor. The proposed work builds heavily on the preliminary EM data of in vitro reconstitution of ASC PYD domain, a common adaptor in most inflammasomes. For the independent phase, Dr. Yin will 1) investigate the assembly mechanisms of NLRP3 inflammasome and elucidate the differential requirements among various inflammasomes (Aim 2), and 2) assay structure-based stabilized helical peptides and screen chemical libraries for small molecules that can perturb inflammasome assembly, in the hope to find druggable interfaces and tool compounds with potential therapeutic values (Aim 3). RELEVANCE: Inflammation is a self-protection process that needs to be tightly regulated. Understanding inflammasome activation and regulation will provide new avenues for treatment of many metabolic diseases, e.g. diabetes and various cancers, as well as hereditary autoinflammatory diseases.